Paenibacillus Strains with Reduced EPS Viscosity

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Solution Overview

Problem

Existing methods to reduce exopolysaccharide (EPS) production in Paenibacillus strains during large-scale fermentation are either too drastic, affecting the organism's metabolism, or do not effectively reduce EPS production, especially in the presence of high glucose or saccharose concentrations.

Innovation Solution

Developing Paenibacillus strains with reduced activity of specific enzymes such as PepR, PepH, PepX, ManC, or SacB, which are involved in EPS production, to decrease the viscosity of liquid cultures without severely impacting the organism's metabolic activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If Paenibacillus strains are used to produce exopolysaccharides in liquid culture, then EPS production is enhanced, but viscosity increases excessively making fermentation difficult

Engineering Contradiction:

ImproveEPS productionVSAvoidfermentation process control

Core Design Contradiction:

Quantity of substanceVSEase of operation

Solution Approach 1:

The patent modifies the enzymatic parameters of the Paenibacillus strain by introducing mutations in the sacB gene (levansucrase) and pepR gene (flippase), changing their catalytic activity to reduce EPS production while maintaining viability and other metabolic functions

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent extracts and inactivates specific genes (sacB and pepR) responsible for excessive EPS production, separating the harmful function from the beneficial organism to reduce viscosity while preserving other useful traits

Inventive Principle:

Principle #2Taking out (Extraction)

2Ease of operation

If gene deletions are performed to reduce EPS production, then viscosity decreases, but metabolic activity and organism viability are severely impacted

Engineering Contradiction:

Improveviscosity controlVSAvoidmetabolic activity

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

Instead of complete gene deletion, the patent applies partial inactivation through point mutations in sacB and pepR genes, reducing EPS production to an acceptable level while preserving sufficient enzymatic activity for metabolic functions and organism viability

Inventive Principle:

Principle #16Partial or excessive action

Solution Approach 2:

The patent changes the functional parameters of the genes through mutations, altering enzyme activity levels rather than eliminating them completely, thus achieving a balance between viscosity reduction and metabolic preservation

Inventive Principle:

Principle #35Parameter changes

3Productivity

If EPS production is reduced to improve downstream processing, then manufacturing efficiency increases, but biofilm formation capability is compromised

Engineering Contradiction:

Improvedownstream processing efficiencyVSAvoidbiofilm formation

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent applies local modifications to specific genes (sacB and pepR) involved in EPS production, leaving other genes intact to preserve biofilm formation capabilities and other essential functions while reducing overall EPS viscosity

Inventive Principle:

Principle #3Local quality

Data Source

PatentUS20250145939A1Paenibacillus strains producing low amounts of exopolysaccarides

Publication Date: 2025.05.08 BASF SE

AI summary

Disclosed are Paenibacillus strains comprising a reduced activity of a flippase PepR aflippase PepH, a mannose-1-phosphate guanylyl transferase and/or a levansucrase SacB. These Paenibacillus strains show lower viscosity when grown in liquid culture and agricultural compositions comprising these strains and methods of making and using these strains.